[Sjogren's syndrome treated by Chinese herbal drugs and determination of hermorheologic factors].
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Adult↗
Biomedical subjects
L Tao
Publications and source records attributed to L Tao.
[Study on the relationship among ossification of finger's bone, maxillary and mandibular lengths and body height for Shanghai adolescents aged between 11-15 years old]
OBJECTIVE:To research the intrinsic relationship in the growth and development progress of adolescents and to provide scientific basis for orthodontic diagnosis and treatment.METHODS:Sample included 125 Shanghai adolescents between 11-15 years old, and the relationship among maxillary length, mandibular length, ossification of finger's bone, body height and the age of first coming of menstruation on female was studied.RESULTS:The ossification of finger's bone, the increase of maxilla and mandibular lengths and the body height of Shanghai adolescents between 11-15 years old keep the same pace tendency. Female goes into the young quick growing period 2 years earlier than male, but from about 13 years old the growing speed of male is apparently more quick than that of female. When the ossification of finger's bone is nearly 100%, the body height and the mandibular length of male still possesses great growing capacity. The age of sesamoid bone appearance for both male and female and the age of first coming menstruation for female are getting earlier when comparing with others regions.CONCLUSION: There is a close relationship among ossification of finger's bone, maxilla and mandibular lengths and body height of adolescents. Understanding the intrinsic rule correctly has the guiding significance to early diagnosis and treatment in clinical orthodontics.
Journal Article↗
[A correlate study on the first cervical vertebra and mandibular growth:A comparison of Shanghai females between 12 and 15 years old]
OBJECTIVE:To study the correlation between the cervical vertebra and mandibular growth and to reveal the intrinsic rule of the progress of individual growth, especially the trend of craniofacial development.METHODS:The morphological features of the first cervical vertebra and certain variables of craniofacial in lateral cephalometric radiographs from Shanghai females between 12 and 15 years old were measured and correlated. A set of normal values of the atlas vertebra shape of Shanghai 12 and 15 years old females were established.RESULTS:There was a significant correlation between D1 and AP of the first cervical vertebra and the direction of mandibular growth at the age of 12. But the association between atlas size ratio (AP/D1) and the mandibular growth is more notable than either AP or D1. When the atlas size ratio increased, a reduction of the amount of mandibular horizontal growth was showed, and the mandibular rotated backward; on the contrary, when the atlas size ratio decreased, the amount of mandibular horizontal growth increased and the mandibular rotated forward. CONCLUSION: The first cervical vertebra can be used as an indicator to predict the mandibular growth.
Journal Article↗
Protective mucosal immunity elicited by targeted lymph node immunization with a subunit SIV envelope and core vaccine in macaques.
Prevention of sexually transmitted HIV infection was first investigated in non-human primates by mucosal immunization via the rectal, vaginal or male urethral route. This was compared with subcutaneous targeted iliac lymph node (TILN) and systemic intramuscular immunization in non-human primates. TILN immunization elicited the most consistent mucosal sIgA and IgG antibody response in the rectum, vagina, urine and seminal fluid, as well as in blood. Both mucosal and TILN immunization induced a specific CD4+ T cell proliferative response in the iliac lymph nodes which drain these mucosal surfaces, and in the splenic and circulating T cells. In the next experiment macaques were immunized by the TILN route with SIV gp120 and p27 in alum. Rectal mucosal challenge with SIVmac 32H J5 molecular clone (or cell-free virus) induced total protection in four out of seven macaques, compared with infection in 13 of 14 unimmunized macaques or immunized by other routes (p = 0.025). The remaining three macaques immunized by the TILN route showed either decrease in viral load (> 90%) or transient viraemia, indicating that all seven TILN immunized macaques showed total or partial protection of rectal transmission by SIV (p = 0.001). Protection was associated with significant increase in the iliac lymph nodes IgA antibody secreting cells to p27 (p < 0.02), CD8-suppressor factor inhibiting replication of SIV in CD4+ T cells (p < 0.01) and the chemokines RANTES and MIP-1 beta (p < 0.01). We suggest that administration of gp120 and p27 by the TILN route may elicit protective B and T cell immunity which can significantly prevent rectal transmission of SIV or HIV.
Animals↗
Differential DNA replication origin activities in human normal skin fibroblast and HeLa cell lines.
A modification of the extrusion method for the isolation of nascent DNA from mammalian cells and a PCR-based assay has been used in order to compare the in vivo activities of DNA replication origins in different cell lines. Conventional PCR was firstly applied to detect the chromosomal activities of several known (origins associated with c-myc, hsp70, beta-globin, immunoglobulin mu-chain enhancer) and putative DNA replication origins (autonomously replicating sequences obtained from enriched libraries of human origins of DNA replication from normal and transformed cells) in four human cell lines (HeLa, NSF, WI-38 and SK-MG-1). Then, in nascent DNA samples from normal skin fibroblast (NSF) and HeLa cells, abundance of DNA sequences in the regions of five of these origins was determined by competitive PCR. Our results suggest that autonomously replicating sequences NOA3, S14, S3 and F15 are associated with functional chromosomal origins of replication. Quantitative comparison of origin activities demonstrates that origins associated with c-myc and NOA3 are approximately twice as active in HeLa cells as in NSF cells. The described approach can facilitate the identification of origins which may be differentially active in normal cells and transformed cells or in different cell types.
Cell Line, Transformed↗
Turnover of the acyl phosphates of human and murine prothymosin alpha in vivo.
Prothymosin alpha is a small, highly acidic, abundant, nuclear, mammalian protein which is essential for cell growth. Our laboratory has recently shown that primate prothymosin alpha contains stoichiometric amounts of phosphate on the glutamyl groups of the protein and that in vitro the phosphate undergoes rapid hydrolysis or transfer to a nearby serine residue. Here an assay for the presence of acyl phosphates in vivo has been developed by measuring stable phosphoserine and phosphothreonine in vitro. The assay was used to determine the half-life of the acyl phosphates on prothymosin alpha in vivo by pulse-labeling HeLa cells with [32P]orthophosphate and chasing using three different techniques: permeabilization with digitonin to allow extracellular ATP to equilibrate with the intracellular pool; electroporation in the presence of ATP to reduce the specific activity of [32P]ATP by expansion of the pool; and incubation with inorganic phosphate. Regardless of the method, the phosphate turned over with a half-life of 75-90 min. The ability of cells to phosphorylate old prothymosin alpha molecules was established by demonstrating equivalent labeling of the protein with [32P]orthophosphate in the presence and absence of cycloheximide. The half-life of the acyl phosphates was also studied in resting and growing NIH3T3 cells, with measured values of 30-35 and 70 min, respectively. Our data suggest that the "activity" of prothymosin alpha involves the turnover of its acyl phosphates and that it participates in a function common to all nucleated mammalian cells regardless of whether they are quiescent or undergoing rapid proliferation. This is the first measurement of the stability of protein-bound acyl phosphates in vivo.
3T3 Cells↗
Measurement of antibody-antigen dissociation constants using fast capillary electrophoresis with laser-induced fluorescence detection.
The dissociation constant (Kd) of a monoclonal antibody with fluorescein isothiocyanate (FITC)-labeled insulin and unlabeled insulins from several species were measured using capillary electrophoresis with laser-induced fluorescence detection (CE-LIF). Kd determinations were made by separating free FITC-insulin and its complex with the antibody in equilibrated solutions in 6 s or less. The use of LIF detection allowed quantification of free and bound FITC-insulin in the picomolar range, as is required to measure Kd's below 1 nM. The Kd of FITC-insulin with the antibody was determined to be 0.25 nM by Scatchard analysis. The Kd's of the antibody with unlabeled insulins from several species were obtained by fitting bound over free FITC-insulin as a function of unlabeled insulin concentration data from a series of solutions containing a fixed concentration of FITC-insulin and antibody and variable concentrations of insulin to the expected curve derived from the equilibria and mass balance of the solutions. Kd's for the different insulins were between 0.34 and 0.64 nM.
Animals↗
Comparison of the Ca2+ movement by activation of alpha1-adrenoceptor subtypes in HEK-293 cells.
We studied the Ca2+ movement induced by activation of alpha1A-, alpha1B- and alpha1D-adrenoceptor subtypes in transfected HEK-293 cells with the fura-2 probe. All these alpha1-AR subtypes induced both Ca2+ release and Ca2+ entry. The effect on Ca2+ release in alpha1b transfected HEK-293 cells was bigger than that in alpha1a and alpha1d transfected HEK-293 cells, and the effects on Ca2+ entry were the same in alpha1a, alpha1b and alpha1d transfected HEK-293 cells. The Ca2+ entry was inhibited by 1 mM NiSO4, but not by nifedipine. Cyclopiazonic acid (CPA) produced a biphasic Ca2+ signal response in Ca2+ medium, and only induced a transient response in Ca2+-free medium. After depletion of CPA-sensitive Ca2+ pool by 10 microM CPA in Ca2+-free medium, 10 microM adrenaline (Adr) still transiently increased [Ca2+]i in three different alpha1-adrenoceptor subtype transfected HEK-293 cells. However, after depletion of adrenaline-sensitive Ca2+ pool by 10 microM Adr, CPA transiently elevated [Ca2+]i only in alpha1a and alpha1d transfected HEK-293 cells, not in alpha1b transfected HEK-293 cells. U73122, a phospholipase C (PLC) inhibitor, inhibited both Ca2+ release and Ca2+ entry induced by activation of alpha1A alpha1B and alpha1D subtypes in transfected HEK-293 cells. These results suggest that HEK-293 cell line contains two functionally separate intracellular Ca2+ pools, CPA-sensitive and Adr-sensitive pools. Activation of alpha1B-AR stimulates Ca2+ release from both CPA-sensitive and Adr-sensitive Ca2+ pools. Alpha1A and alpha1D subtypes induce Ca2+ release only from Adr-sensitive Ca2+ pool.
Animals↗
Cerebral astrocytes transport ascorbic acid and dehydroascorbic acid through distinct mechanisms regulated by cyclic AMP.
Cerebral ischemia and trauma lead to rapid increases in cerebral concentrations of cyclic AMP and dehydroascorbic acid (DHAA; oxidized vitamin C), depletion of intracellular ascorbic acid (AA; reduced vitamin C), and formation of reactive astrocytes. We investigated astrocytic transport of AA and DHAA and the effects of cyclic AMP on these transport systems. Primary cultures of astrocytes accumulated millimolar concentrations of intracellular AA when incubated in medium containing either AA or DHAA. AA uptake was Na+-dependent and inhibited by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), whereas DHAA uptake was Na+-independent and DIDS-insensitive. DHAA uptake was inhibited by cytochalasin B, D-glucose, and glucose analogues specific for facilitative hexose transporters. Once inside the cells, DHAA was reduced to AA. DHAA reduction greatly decreased astrocytic glutathione concentration. However, experiments with astrocytes that had been previously depleted of glutathione showed that DHAA reduction does not require physiological concentrations of glutathione. Astrocyte cultures were treated with a permeant analogue of cyclic AMP or forskolin, an activator of adenylyl cyclase, to induce cellular differentiation and thus provide in vitro models of reactive astrocytes. Cyclic AMP stimulated uptake of AA, DHAA, and 2-deoxyglucose. The effects of cyclic AMP required at least 12 h and were inhibited by cycloheximide, consistent with a requirement for de novo protein synthesis. Uptake and reduction of DHAA by astrocytes may be a recycling pathway that contributes to brain AA homeostasis. These results also indicate a role for cyclic AMP in accelerating the clearance and detoxification of DHAA in the brain.
Animals↗
Chemopreventive agents-induced regression of azoxymethane-induced aberrant crypt foci with the recovery of hexosaminidase activity.
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Animals↗
Metabolic regulation of lrp gene expression in Escherichia coli K-12.
Expression of the lrp gene is regulated in part by the nutrients available to the cell, and is decreased in rich medium, in glucose minimal media enriched with amino acids, and in minimal medium with alternative carbon sources, such as acetate and succinate. When Lrp production is increased in a given medium, expression of its target genes is also increased. However, when the medium is changed from glucose to acetate, the response of the target genes is governed by many factors.
Amino Acids↗
Role of extracellular signal-regulated kinase and PKC alpha in cytosolic PLA2 activation by bradykinin in MDCK-D1 cells.
The actions of bradykinin (BK) in Madin-Darby canine kidney (MDCK) and other cell types involve formation of arachidonic acid (AA) and AA products by as-yet-undefined mechanisms. We found that BK promoted AA release and an increase in phospholipase A2 (PLA2) activity in subsequently prepared MDCK-D1 cell lysates, both of which were Ca2+ dependent and were inhibited by the 85-kDa cytosolic PLA2 (cPLA2) inhibitor arachidonyl trifluoromethyl ketone. In addition, BK treatment of cells led to increased PLA2 activity of cPLA2 immunoprecipitated from lysates. Thus BK receptors mediate AA release via cPLA2 in MDCK-D1 cells. The BK-promoted increase of cPLA2 activity was reversed by treatment of cell lysates with potato acid phosphatase, implying that phosphorylation underlies the activation of cPLA2. However, extracellular signal-regulated kinase (ERK) appeared not to be responsible for this phosphorylation, because treatment of cells with BK (in contrast with the results obtained with epinephrine and phorbol ester) caused neither enzyme activation nor phosphorylation (as judged by molecular mass shift) of this kinase. Although the alpha isoform of protein kinase C (PKC alpha) is responsible for AA release promoted by phorbol ester treatment of MDCK-D1 cells (C. Godson, K.S. Bell, and P.A. Insel. [corrected] J. Biol. Chem. 268: 11946-11950, 1993), neither treatment of cells with the PKC alpha-selective inhibitor GF109203X nor transfection of cells with PKC alpha antisense cDNA altered BK-mediated AA release. We conclude that PKC alpha is unlikely to play an important role in the regulation of cPLA2 by BK receptors in MDCK-D1 cells. The tyrosine kinase inhibitor herbimycin A, on the other hand, inhibited both BK-promoted AA release in intact cells and cPLA2 activation in cell lysates, suggesting the involvement of tyrosine kinase in the regulation of this lipase by BK receptors. Taken together, these data suggest that BK receptors in MDCK-D1 cells regulate cPLA2 via phosphorylation mediated by kinases other than ERK and PKC alpha.
Animals↗
Interleukin 1 in experimental proliferative vitreoretinopathy.
Since proliferative vitreoretinopathy (PVR) is associated with an inflammatory reaction and interleukin-1 (IL-1) initiates a cascade of inflammatory mediators, we determined the intravitreal levels of IL-1 in experimental PVR. In the blood-trauma model of PVR in rabbits, we measured the intravitreal levels of IL-1 alpha and IL-1 beta using a radioimmunoassay and performed immunohistochemical staining of cells from the vitreous. We found that on day 1 polymorphonuclear and from day 2 on mononuclear inflammatory cells predominated in the vitreous. IL-1 alpha- and IL-1 beta-positive cells with a large cytoplasm were observed on days 4 and 8. However, the IL-1 alpha and IL-1 beta concentrations in the vitreous did not exceed the radioimmunoassay sensitivity limit 8 h, 1, 2, 4 and 8 days after trauma, while in 2 control rabbits injected with additional endotoxins we found 5.6 and 8 ng/ml IL-1 beta. We conclude that despite IL-1-positive cells, presumably macrophages, PVR is not associated with high levels of IL-1. No conclusions concerning the low levels of IL-1 alpha and IL-beta can be drawn due to the performance of the RIA.
Animals↗
Host-pathogen interactions in bacterial endocarditis: streptococcal virulence in the host.
To identify streptococcal genes that are expressed during experimental endocarditis, we developed a promoter-less dual reporter gene-fusion (amy, cat) plasmid, pAK36. Chromosomal DNA from S. gordonii V288 was digested with Sau3A1. The resulting fragments were ligated into pAK36. Following transformation into S. gordonii, the library of random gene fusion clones was inoculated into a rabbit to induce experimental endocarditis. Chloramphenicol treatment effected positive selection. Upon euthanization of the rabbits, the valvular vegetations were excised in a sterile field. Surviving clones were isolated and screened in vitro for chloramphenicol sensitivity and negative amylase activity. From the 48 randomly picked, double-negative clones, DNA was isolated and analyzed by Southern hybridization with labeled pAK36 probe. Different insertion patterns were identified, suggesting that no fewer than 13 S. gordonii genes were induced. Therefore, S. gordonii genes are induced during experimental endocarditis, which may contribute to virulence.
Animals↗
Inhibitory effects of nifedipine on DNA and protein synthesis in cultured cardiac nonmyocytes of neonatal rats.
AIM: To observe the inhibitory effects of nifedipine (Nif) on cardiac nonmyocytes growth and proliferation. METHODS: Using nonmyocytes in culture as a model, [3H]thymidine and [3H]leucine incorporation were measured. RESULTS: There was a significant decrease in cell number and in total cellular protein after 72-h exposure to Nif 1 mumol.L-1 in the presence of angiotensin II (Ang II). A 48-h exposure to 1, 10, 100, 1000 nmol.L-1 of Ang II caused a 19%, 35%, 46%, 48% increase in protein synthesis and 27%, 46%, 56%, 57% increase in DNA synthesis. Nif 1, 3, and 10 mumol.L-1 were able to reduce the Ang II 100 nmol.L-1-induced increase of protein synthesis and DNA synthesis. CONCLUSION: Nif had a direct inhibitory action on the growth of nonmyocytes, which was related to the regression of cardical hypertrophy.
Angiotensin II↗
Loss of heterozygosity on chromosome 6 in dichloroacetic acid and trichloroacetic acid-induced liver tumors in female B6C3F1 mice.
Dichloroacetic acid (DCA) and trichloroacetic acid (TCA) are major metabolites of tetrachloroethylene (PCE) and trichloroethylene (TCE) and are found in chlorinated drinking water. All four chlorinated compounds are liver carcinogens in B6C3F1 mice. It has previously been reported that approximately 20% of hepatic tumors induced by PCE exhibited loss of heterozygosity (LOH) on chromosome 6, suggesting the presence of a tumor suppressor gene. In the current investigation, we determined whether TCA or DCA also induced LOH on chromosome 6. Liver tumors were initiated in 15 day old female B6C3F1 mice with N-methyl-N-nitrosourea (MNU) and promoted with 20 mmol/l DCA or TCA in their drinking water. Twenty-four and thirty-seven liver tumors promoted by DCA and TCA, respectively, were examined for LOH using 4 polymorphic loci on chromosome 6. Ten of 37 (27%) tumors (7 of 27 carcinomas and 3 of 10 adenomas) promoted by TCA exhibited LOH at least for two loci on chromosome 6. All 10 tumors that exhibited LOH, lost the C57BL/6J allele at both the D6mit9 loci, while two also lost at least one of the C3H/HeJ alleles. No LOH on chromosome 6 was observed in the 24 liver tumors promoted by DCA. The LOH on chromosome 6 in TCA but not in DCA-promoted tumors supports it as an active metabolite of PCE and demonstrates different pathogenesis at least for some of the DCA and TCA-promoted liver cancer.
Animals↗
On-line competitive immunoassay for insulin based on capillary electrophoresis with laser-induced fluorescence detection.
An on-line competitive immunoassay for insulin has been developed and applied to monitoring insulin concentration in a flowing stream. In the assay, solutions of fluorescein-labeled insulin (FITC-insulin), monoclonal anti-insulin, and sample containing insulin are pumped into a cross where they begin to mix. The mixture flows through a fused silica reactor capillary to a flow-gated interface. During transfer to the interface, insulin and FITC-insulin compete to form a complex with the antibody. At the interface, a plug of the mixture is injected into a separation capillary, where the bound and free FITC-insulin are separated and detected by capillary electrophoresis with laser-induced fluorescence detection. The amount of bound FITC-insulin, amount of free FITC-insulin, or bound/free ratio can be used to quantify insulin concentration. Typical relative standard deviations of bound over free ratio are 5%. The detection limit of the immunoassay in the on-line mode is < 0.3 nM. Each separation requires as little as 3 s, and over 1600 consecutive assays can be acquired with no need to rinse the separation capillary. Thus, the system can be used to monitor insulin in a flowing stream for flow injection analysis or for sensor-like monitoring. Dilution and zone broadening during transfer of sample to the interface limit the response time of the on-line system to about 25 s. As a demonstration of the on-line immunoassay, the insulin content of single islets of Langerhans was determined by flow injection analysis.
Electrophoresis, Capillary↗
Diffusion of albumins in rat cortical slices and relevance to volume transmission.
The apparent diffusion coefficient, D*, was measured in rat cortical slices and compared to the free diffusion coefficient, D, for three negatively charged proteins, lactalbumin (mol. wt = 14,500), ovalbumin (45,000) and bovine serum albumin (66,000). The temporal evolution of the spatial distribution of albumin molecules labeled with the Texas Red fluorophore was determined using integrative optical imaging at intervals after a brief pressure injection from a micropipette in slices of adult rat cerebral cortex and dilute agarose gel. Diffusion coefficients were obtained by fitting appropriate equations to the data. In slices at 34 degrees C, the values of D* (10(-7) cm2/s, mean +/- S.E.M.) for lactalbumin, ovalbumin and bovine serum albumin were 2.37 +/- 0.10, 1.60 +/- 0.08 and 1.63 +/- 0.07, respectively. In agarose gel, values of D (10(-7) cm2/s) were 11.87 +/- 0.20, 10.02 +/- 0.25 and 8.29 +/- 0.17, respectively. From these data the tortuosity factors, (D/D*)0.5, were calculated, with 2.24 obtained for lactalbumin, 2.50 for ovalbumin and 2.26 for bovine serum albumin. Previous optical measurements using dextrans with mol. wts of 40,000 and 70,000 gave tortuosities of 2.16 and 2.25, but in contrast previous determinations with ion-selective microelectrodes using the small cation tetramethylammonium (mol. wt = 74.1) give tortuosities of about 1.6. The results show that proteins as large as bovine serum albumin diffuse through brain extracellular space but are more hindered than smaller molecules. A simple model compared the differences in diffusion properties of bovine serum albumin, dopamine and nitric oxide in brain tissue and discussed the implications for volume transmission of chemical information between cells. The results are also relevant to the behavior of diffusible factors in brain development and the delivery of therapeutic agents.
Albumins↗